| ObjectiveThe infusion of antitumor drugs in urinary bladder cavity is one of the important methods in bladder carcinoma treatment. But the effect of this method is not satisfactory overall. More dose of drugs may improve the effect, but most of patients couldnt endure the side effects. Reasonable combination of drugs may bring about synergistic effect, which can not only improve the effect but also decrease the drug concentration.Fas is an transmembrane protein of type I , which may activate Caspase -1,3 through a series of complicated mechanism when combined with its natural ligand or antibody. After that the target cells will apoptosis. Bcl -2 is an inhibitory gene in apoptosis. Studies showed that mitomycin(MMC) may down regulate the gene expression of Bl -2. Whether MMC and anti - Fas mAb may bring a-bout synergistic effect to the cell line T24, which is positive in Fas? It hasn' t been reported at this point by now.The method of Chou - Talalay association index (i. e. median - effect principle) may quantitively analysis the synergism, admoveantur and antagonistic effect. In abroad, this method has been used by more and more scholar, while in our country it hasnt been acknowledged generally. In the field of urinary surgery , on report on the method of Chou - Talalay association index has been published. Here, we study the synergistic effect between MMC and anti - Fas mAb in human bladder carcinoma T24 cells with the method of Chou - Talalay associ-ation index.Materials and methods1. MaterialsThe cell line is human bladder carcinoma T24. The drugs we used included MMC, anti - Fas mAb, MTT, medium of DMEM, new -born calf serum, etc. Main experiment facilities included cell culture incubator, super clean bench, inverted microscope, automatic enzyme detection implement etc.2. Methods2.1 The effect of drugs by MTTWhen the drugs were used alone, the final concentrations of MMC: 11. 964, 23.928, 47.856, 95.711, 191. 422 μmol/L; anti-Fas mAb: 0.278 × 10-3, 0.556 ×10-3, 1. 111 × 10-3, 2.222 × 10-3, 4.444 × 10-3 μmol/L. When they were used together, the ratio is 1:1. After 12 hours of drug disposal, MTT solution was added. Then the cells were incubated for4 hours. After that, the supernatant was removed. DMSO was to dissolve the crystal. Then the ab-sorbance A at 570 nm was determined by automatic detective meter. The inhibitory rate could be calculated with the consequence.2. 2 Consequence assessmentAccording to median - effect equation fa/fu = ( D/Dm) m, (fa is effect, fu = 1 - fa, D is drug concentration, Dm is median - effect concentration, i. e. the concentration when the effect is 50% , m is slope rate) , we could calculate the concentration [ D = Dm ( fa/fu)1/m ] whether one drug was used alone or combined with another, reaching any effect. Then we may gain the combination index(CI) when the two drugs were combinated; CI = (D) 1/( Dx)1+ (D)2/ (Dx)2 +a (D)1(D)2/(Dx)1(Dx)2. (D)1 and (D)2 are the concentrations when the two drugs were combined, reaching a effect X. (Dx)1 and (Dx)2 are the concentrations when the two drugs were used alone, reaching a effect X. When the two drugs' mechanism are different,a = 1. Otherwiseα =0. CI < 1 means synergism. CI = 1 means admoveantur and CI > 1 means antagonism. |
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